紫葳科植物猫爪藤阿克替苷的提取富集工艺

Extraction and Enrichment of Acteoside from Macfadyena unguis-cati(Bignoniaceae)

建立UPLC测定猫爪藤Macfadyena unguis-cati阿克替苷含量的方法,采用单因素实验和正交试验优化阿克替苷提取工艺,通过静态、动态吸附与解吸实验筛选适合富集阿克替苷的大孔树脂,并对富集工艺进行研究。UPLC条件:色谱柱HSST3柱(2.1×100 mm,1.8μm);流动相0.1%甲酸(A)-乙腈(B);梯度90%A 5 min,75%A 7.5 min,30%A 9 min,90%A;检测波长310 nm;柱温25℃;流速0.3 mL·min^(-1);保留时间5.21 min。在UPLC条件下阿克替苷在0.25~100 ng进样范围内峰面积与进样量呈良好的线性关系,回归方程为Y=89.002X-4.8275,R=0.9999。单因素实验显示,70%乙醇为最佳提取溶剂。正交试验确定最佳提取条件为70%乙醇,料液比1∶10,提取时间12 h。静态、动态吸附和解吸实验确定HP-20大孔树脂为最佳富集材料。富集工艺为4倍体积15%乙醇洗脱除杂后收集4倍体积40%乙醇洗脱的部分。

A UPLC method for the determination of acetoside in Macfadyena unguis-cati was established. Single factor and orthogonal experiments for acetoside extraction from M. unguis-cati were designed. Macroporous resins were screened for the enrichment of acetoside and the elution procession was determinated. The UPLC conditions： HSST3 column （2.1 ~ 100 mm, 1.8 ~tm）, mobile phase 0.1% HCOOH（A）-CH3CN（B）, DAD detector at 310 nm, flow rate 0.3 mL.min-1, column temperature 25 ~C, elution process： 90%A-5 min, 75%A-7.5 min, 30%A-9 min, 90%A. The results showed that the calibration curves of acetoside was in a good linearity over the range of 0.25--100 ng under the experiment condition and the regression equation was Y=89.002X-4.8275 （R=0.9999）. The best extraction solvent was 70% ethanol according single factor experiment and the optimum extraction conditions were 70% ethanol, solid-liquid ratio of 1 ＂ 10 and 12 h extraction time through orthogonal experiments. The most suitable macroporous resin for acetoside enrichment was HP-20 through static and dynamic adsorption and desorption experiments. The enrichment process by HP-20 resin was gathering 4 BV of the 40% ethanol elute after 4 BV of elution by 15% ethanol.